Subtracted differential display: genes with amphetamine-altered expression patterns include calcineurin.
Genes whose expression changes with administration of abused substances provide candidate biochemical mechanisms for drug-induced long-term brain changes. To identify such genes, and to avoid the false-positive results frequently obtained from differential display PCR, we applied a subtracted differential display (SDD) approach. We subtracted single-stranded cDNA prepared from drug-treated animals with excess mRNA from saline-treated animals, and visa versa, prior to differential display amplifications. Two of initial amphetamine-regulated cDNAs identified in this fashion encoded calcineurin A, a neuron-specific protein phosphatase catalytic subunit whose striatal expression was upregulated ca. 1.5-fold. SDD may enhance the utility of differential display approaches to identifying regulated genes in tissues in which mRNA complexities are high.
Wang
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Molecular Neurobiology Branch, Intramural Research Program, National Institute on Drug Abuse, Box 5180, Baltimore, MD 21224, USA. xwang@irp.nida.nih.gov
Uhl
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D000818:Animals; D001921:Brain / Q000187:drug effects / Q000201:enzymology*; D019703:Calcineurin / Q000096:biosynthesis*; D003001:Cloning, Molecular; D003342:Corpus Striatum / Q000201:enzymology; D018076:DNA, Complementary; D003913:Dextroamphetamine / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D008297:Male; D009714:Nucleus Accumbens / Q000201:enzymology; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014158:Transcription, Genetic / Q000187:drug effects*
D018076:DNA, Complementary; D012333:RNA, Messenger; D019703:Calcineurin; D003913:Dextroamphetamine